, Volume 41, Issue 4, pp 1334–1348 | Cite as

A Novel Tetrasubstituted Imidazole as a Prototype for the Development of Anti-inflammatory Drugs

  • Marcus Vinicius P. S. Nascimento
  • Antonio C. M. Munhoz
  • Lais C. Theindl
  • Eduarda Talita B. Mohr
  • Najla Saleh
  • Eduardo B. Parisotto
  • Thaís A. Rossa
  • Ariane Zamoner
  • Tania B. Creczynski-Pasa
  • Fabíola B. Filippin-Monteiro
  • Marcus M. Sá
  • Eduardo Monguilhott Dalmarco


Although inflammation is a biological phenomenon that exists to protect the host against infections and/or related problems, its unceasing activation results in the aggravation of several medical conditions. Imidazoles, whether natural or synthetic, are molecules related to a broad spectrum of biological effects, including anti-inflammatory properties. In this study, we screened eight novel small molecules of the imidazole class synthesized by our research group for their in vitro anti-inflammatory activity. The effect of the selected molecules was confirmed in an in vivo inflammatory model. We also analyzed whether the effects were caused by inhibition of nuclear factor kappa B (NF-κB) transcription factor transmigration. Of the eight imidazoles tested, methyl 1-allyl-2-(4-fluorophenyl)-5-phenyl-1H-imidazole-4-acetate (8) inhibited nitric oxide metabolites and pro-inflammatory cytokine (TNF-α, IL-6, and IL-1β) secretion in J774 macrophages stimulated with LPS. It also attenuated leukocyte migration and exudate formation in the pleural cavity of mice challenged with carrageenan. Furthermore, imidazole 8 reverted the oxidative stress pattern triggered by carrageenan in the pleural cavity by diminishing myeloperoxidase, superoxide dismutase, catalase, and glutathione S-transferase activities and reducing the production of nitric oxide metabolites and thiobarbituric acid-reactive substances. Finally, these effects can be attributed, at least in part, to the ability of this compound to prevent NF-κB transmigration. In this context, our results demonstrate that imidazole 8 has promising potential as a prototype for the development of a new anti-inflammatory drug to treat inflammatory conditions in which NF-κB and oxidative stress play a prominent role.

Graphical Abstract


Inflammation imidazole J774 pleurisy oxidative stress NF-κB 



The authors are grateful to Fiona Robson for her assistance in proofreading and correcting the English language of this text.


This study was granted and financially supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (Grant number, 483713/2012-0) and fellowships from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical Approval

All procedures in studies involving animals were performed in accordance with the ethical standards of the institution (CEUA/UFSC Protocol PP00965).

Supplementary material

10753_2018_782_MOESM1_ESM.pdf (406 kb)
ESM 1 Supplementary materials on the general procedure for the synthesis of imidazoles 1–8, including 1H and 13C NMR spectra. (PDF 406 kb)


  1. 1.
    Granger, D.N., and E. Senchenkova. 2010. Colloquium series on integrated systems physiology: from molecule to function to disease. In Inflammation and the microcirculation. San Rafael: Morgan & Claypool Life Sciences.Google Scholar
  2. 2.
    Biswas, S.K. 2016. Does the interdependence between oxidative stress and inflammation explain the antioxidant paradox? Oxidative Medicine and Cellular Longevity 2016: 1–9. Scholar
  3. 3.
    Schmid-Schonbein, G.W. 2006. Analysis of inflammation. Annual Review of Biomedical Engineering 8: 93–131. Scholar
  4. 4.
    Nathan, C., and A. Ding. 2010. Nonresolving inflammation. Cell 140: 871–882. Scholar
  5. 5.
    Samanen, J. 2013. Similarities and differences in the discovery and use of biopharmaceuticals and small-molecule chemotherapeutics. In Introduction to Biological and Small Molecule Drug Research and Development, eds. C. Ganellin, Roy Jefferis, Stanley Roberts, 161–203. Oxford: Elsevier.CrossRefGoogle Scholar
  6. 6.
    Hanke, T., D. Merk, D. Steinhilber, G. Geisslinger, and M. Schubert-Zsilavecz. 2016. Small molecules with anti-inflammatory properties in clinical development. Pharmacology & Therapeutics 157: 163–187. Scholar
  7. 7.
    Zhang, L., X.M. Peng, G.L.V. Damu, R.X. Geng, and C.H. Zhou. 2014. Comprehensive review in current developments of imidazole-based medicinal chemistry. Medicinal Research Reviews 34: 340–437. Scholar
  8. 8.
    Ahmed, S.A., R.M. Jr Gogal, and J.E. Walsh. 1994. A new rapid and simple non-radioactive assay to monitor and determine the proliferation of lymphocytes: an alternative to [3H]thymidine incorporation assay. Journal of Immunological Methods 170: 211–224.CrossRefPubMedGoogle Scholar
  9. 9.
    Green, L.C., D.A. Wagner, J. Glogowski, P.L. Skipper, J.S. Wishnok, and S.R. Tannenbaum. 1982. Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Analytical Biochemistry 126: 131–138. Scholar
  10. 10.
    Flecknell, P. 2002. Replacement, reduction and refinement. ALTEX 19: 73–78. PubMedGoogle Scholar
  11. 11.
    Dalmarco, E.M., T.S. Fröde, and Y.S. Medeiros. 2004. Additional evidence of acute anti-inflammatory effects of cyclosporin a in a murine model of pleurisy. Transplant Immunology 12: 151–157. Scholar
  12. 12.
    Lowry, O.H., N.J. Rosebrough, A.L. Farr, and R.J. Randall. 1951. Protein measurement with the folin phenol reagent. The Journal of Biological Chemistry 193: 265–275.PubMedGoogle Scholar
  13. 13.
    Bradley, P.P., D.A. Priebat, R.D. Christensen, and G. Rothstein. 1982. Measurement of cutaneous inflammation: estimation of neutrophil content with an enzyme marker. Journal of Investigative Dermatology 78: 206–209.CrossRefPubMedGoogle Scholar
  14. 14.
    Miranda, K.M., M.G. Espey, and D.A. Wink. 2001. A rapid, simple spectrophotometric method for simultaneous detection of nitrate and nitrite. Nitric Oxide 5: 62–71. Scholar
  15. 15.
    Misra, H.P., and I. Fridovich. 1972. The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. The Journal of Biological Chemistry 247: 3170–3175.PubMedGoogle Scholar
  16. 16.
    Boveris, A., C.G. Fraga, A.I. Varsavsky, and O.R. Koch. 1983. Increased chemiluminescence and superoxide production in the liver of chronically ethanol-treated rats. Archives of Biochemistry and Biophysics 227: 534–541. Scholar
  17. 17.
    Aebi, H. 1984. Catalase in vitro. Methods in Enzymology 105: 121–126.CrossRefPubMedGoogle Scholar
  18. 18.
    Habig, W.H., M.J. Pabst, and W.B. Jakoby. 1974. Glutathione S transferases. The first enzymatic step in mercapturic acid formation. Journal of Biological Chemistry 249: 7130–7139. PubMedGoogle Scholar
  19. 19.
    Bird, R.P., and H.H. Draper. 1984. Comparative studies on different methods of malonaldehyde determination. Methods in Enzymology 105: 299–305.CrossRefPubMedGoogle Scholar
  20. 20.
    Goldust, M., E. Rezaee, S. Masoudnia, and R. Raghifar. 2013. Clinical study of sertaconazole 2% cream vs. hydrocortisone 1% cream in the treatment of seborrheic dermatitis. Annals of Parasitology 59: 119–123.PubMedGoogle Scholar
  21. 21.
    Umarani, N., K. Ilango, G. Garg, K. Bompada, and V. Hemalatha. 2011. Exploring the effects of newer three component aminobenzylated reactions of triphenyl imidazole motif as potent antimicrobial and anti-inflammatory agents. International Journal of Pharmacy and Pharmaceutical Sciences 3: 62–65.Google Scholar
  22. 22.
    Xu, N., M. Hossain, and L. Liu. 2013. Pharmacological inhibition of p38 mitogen-activated protein kinases affects KC/CXCL1-induced intraluminal crawling, transendothelial migration, and chemotaxis of neutrophils in vivo. Mediators of Inflammation 290565: 1–10. Scholar
  23. 23.
    Cuzzocrea, S., E. Mazzon, G. Calabro, L. Dugo, A. De Sarro, F.A. van De Loo, and A.P. Caputi. 2000. Inducible nitric oxide synthase-knockout mice exhibit resistance to pleurisy and lung injury caused by carrageenan. American Journal of Respiratory and Critical Care Medicine 162: 1859–1866. Scholar
  24. 24.
    Luz, A.B.G., C.H.B. Da Silva, M.V.P.S. Nascimento, B.M.C. Facchin, B. Baratto, T.S. Fröde, F.H. Reginatto, and E.M. Dalmarco. 2016. The anti-inflammatory effect of Ilex paraguariensis A. St. Hil (Mate) in a murine model of pleurisy. International Immunopharmacology 36: 165–172. Scholar
  25. 25.
    Dos Reis, G.O., G. Vicente, F.K. de Carvalho, M. Heller, G.A. Micke, M.G. Pizzolatti, and T.S. Frode. 2014. Croton antisyphiliticus Mart. attenuates the inflammatory response to carrageenan-induced pleurisy in mice. Inflammopharmacology 22: 115–126. Scholar
  26. 26.
    Förstermann, U., and W.C. Sessa. 2012. Nitric oxide synthases: regulation and function. European Heart Journal 33: 829–837. Scholar
  27. 27.
    Wolff, D.J., G.A. Datto, R.A. Samatovicz, and R.A. Tempsick. 1993. Calmodulin-dependent nitric-oxide synthase. Mechanism of inhibition by imidazole and phenylimidazoles. Journal of Biological Chemistry 268: 9425–9429.PubMedGoogle Scholar
  28. 28.
    Nussbaum, C., A. Klinke, M. Adam, S. Baldus, and M. Sperandio. 2012. Myeloperoxidase: a leukocyte-derived protagonist of inflammation and cardiovascular disease. Antioxidants & Redox Signaling 18: 121003062117006. Scholar
  29. 29.
    Odobasic, D., Y. Yang, R.C.M. Muljadi, K.M. O’Sullivan, W. Kao, M. Smith, E.F. Morand, and S.R. Holdsworth. 2014. Endogenous myeloperoxidase is a mediator of joint inflammation and damage in experimental arthritis. Arthritis & Rheumatology 66: 907–917. Scholar
  30. 30.
    Winterbourn, C.C., and A.J. Kettle. 2013. Redox reactions and microbial killing in the neutrophil phagosome. Antioxidants & Redox Signaling 18: 642–660. Scholar
  31. 31.
    Silva, V.G., R.O. Silva, S.R.B. Damasceno, N.S. Carvalho, R.S. Prudeîncio, K.S. Aragão, M.A. Guimarães, et al. 2013. Anti-inflammatory and antinociceptive activity of epiisopiloturine, an imidazole alkaloid isolated from pilocarpus microphyllus. Journal of Natural Products 76: 1071–1077. Scholar
  32. 32.
    Sánchez-Gómez, F.J., B. Díez-Dacal, E. García-Martín, J.A.G. Agúndez, M.A. Pajares, and D. Pérez-Sala. 2016. Detoxifying enzymes at the cross-roads of inflammation, oxidative stress, and drug hypersensitivity: role of glutathione transferase P1-1 and aldose reductase. Frontiers in Pharmacology 7: 1–9. Scholar
  33. 33.
    Halici, Z., G.O. Dengiz, F. Odabasoglu, H. Suleyman, E. Cadirci, and M. Halici. 2007. Amiodarone has anti-inflammatory and anti-oxidative properties: an experimental study in rats with carrageenan-induced paw edema. European Journal of Pharmacology 566: 215–221. Scholar
  34. 34.
    Mittal, M., M.R. Siddiqui, K. Tran, S.P. Reddy, and A.B. Malik. 2014. Reactive oxygen species in inflammation and tissue injury. Antioxidants & Redox Signaling 20: 1126–1167. Scholar
  35. 35.
    Dalmarco, E.M., P. Budni, E.B. Parisotto, D.W. Filho, and T.S. Frode. 2009. Antioxidant effects of mycophenolate mofetil in a murine pleurisy model. Transplant Immunology 22: 12–17. Scholar
  36. 36.
    Thomas, S.R., P.K. Witting, and G.R. Drummond. 2008. Redox control of endothelial function and dysfunction: molecular mechanisms and therapeutic opportunities. Antioxidants & Redox Signaling 10: 1713–1766. Scholar
  37. 37.
    Sorrenti, V., L. Salerno, C. Di Giacomo, R. Acquaviva, M.A. Siracusa, and A. Vanella. 2006. Imidazole derivatives as antioxidants and selective inhibitors of nNOS. Nitric Oxide—Biology and Chemistry 14: 45–50. CrossRefPubMedGoogle Scholar
  38. 38.
    Becher, B., S. Spath, and J. Goverman. 2016. Cytokine networks in neuroinflammation. Nature Reviews Immunology 17: 49–59. Scholar
  39. 39.
    Navarro-Millán, I., J.A. Singh, and J.R. Curtis. 2012. Systematic review of tocilizumab for rheumatoid arthritis: a new biologic agent targeting the interleukin-6 receptor. Clinical Therapeutics 34: 788–802.e3. Scholar
  40. 40.
    Pazyar, N., A. Feily, and R. Yaghoobi. 2012. An overview of interleukin-1 receptor antagonist, anakinra, in the treatment of cutaneous diseases. Current Clinical Pharmacology 7: 271–275. Scholar
  41. 41.
    Scott, L.J. 2014. Etanercept: a review of its use in autoimmune inflammatory diseases. Drugs 74: 1379–1410. Scholar
  42. 42.
    Laufer, S.A., W. Zimmermann, and K.J. Ruff. 2004. Tetrasubstituted imidazole inhibitors of cytokine release: probing substituents in the N-1 position. Journal of Medicinal Chemistry 47: 6311–6325. Scholar
  43. 43.
    Zhou, W.D., H.M. Yang, Q. Wang, D.Y. Su, F.A. Liu, M. Zhao, Q.H. Chen, and Q.X. Chen. 2010. SB203580, a p38 mitogen-activated protein kinase inhibitor, suppresses the development of endometriosis by down-regulating proinflammatory cytokines and proteolytic factors in a mouse model. Human Reproduction 25: 3110–3116. Scholar
  44. 44.
    Christian, F., E. Smith, and R. Carmody. 2016. The regulation of NF-κB subunits by phosphorylation. Cell 5: 12. Scholar
  45. 45.
    Lawrence, T. 2009. The nuclear factor NF-κB pathway in inflammation. About Cold Spring Harbor Perspectives in Biology 1: 1–10. Scholar
  46. 46.
    Cushing, T.D., V. Baichwal, K. Berry, R. Billedeau, V. Bordunov, C. Broka, M. Cardozo, P. Cheng, D. Clark, S. Dalrymple, M. DeGraffenreid, A. Gill, X. Hao, R.C. Hawley, X. He, J.C. Jaen, S.S. Labadie, M. Labelle, C. Lehel, P.P. Lu, J. McIntosh, S. Miao, C. Parast, Y. Shin, E.B. Sjogren, M.L. Smith, F.X. Talamas, G. Tonn, K.M. Walker, N.P.C. Walker, H. Wesche, C. Whitehead, M. Wright, and M.F. Browner. 2011. A novel series of IKKβ inhibitors part I: initial SAR studies of a HTS hit. Bioorganic and Medicinal Chemistry Letters 21: 417–422. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Marcus Vinicius P. S. Nascimento
    • 1
  • Antonio C. M. Munhoz
    • 1
  • Lais C. Theindl
    • 2
  • Eduarda Talita B. Mohr
    • 1
  • Najla Saleh
    • 1
  • Eduardo B. Parisotto
    • 3
  • Thaís A. Rossa
    • 4
  • Ariane Zamoner
    • 3
  • Tania B. Creczynski-Pasa
    • 5
  • Fabíola B. Filippin-Monteiro
    • 2
  • Marcus M. Sá
    • 4
  • Eduardo Monguilhott Dalmarco
    • 2
  1. 1.Postgraduation Program in PharmacyFederal University of Santa CatarinaFlorianopolisBrazil
  2. 2.Department of Clinical Analysis, Centre of Health SciencesFederal University of Santa CatarinaFlorianópolisBrazil
  3. 3.Department of BiochemistryFederal University of Santa CatarinaFlorianopolisBrazil
  4. 4.Department of ChemistryFederal University of Santa CatarinaFlorianopolisBrazil
  5. 5.Department of Pharmaceutical SciencesFederal University of Santa CatarinaFlorianopolisBrazil

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